Urea inhibits Na-K-2Cl cotransport in medullary thick ascending limb cells

We examined the effect of physiological concentrations of urea (100-500 mM) on Na-K-2Cl cotransport in cultured cells from mouse medullary thick ascending limb (mTAL). Urea acutely inhibited bumetanide-sensitive K influx in mTAL cells in a concentration-dependent fashion, with a statistically significant inhibition (19%) at 100 mM and 86% inhibition at 500 mM. The effect of urea was entirely reversible and was blocked by prior treatment with okadaic acid, a phosphatase inhibitor, suggesting that urea exerts its action upstream of the phosphorylation-dephosphorylation step. Cell volume was unchanged in the presence of 500 mM urea. The number of [3H]bumetanide binding sites, a measure of the number of functioning cotransporter sites, was decreased in the presence of urea, and the decrease in bumetanide binding was proportional to the decrease in bumetanide-sensitive K influx. Urea also stimulated the Ba-sensitive swelling-activated K efflux from mTAL cells. Thus urea, in concentrations that prevail in the renal medulla, alters ion transport in mTAL cells.